EP1648607B1 - Stack of supports, in particular for cryopreservation of biological samples - Google Patents
Stack of supports, in particular for cryopreservation of biological samples Download PDFInfo
- Publication number
- EP1648607B1 EP1648607B1 EP04741090A EP04741090A EP1648607B1 EP 1648607 B1 EP1648607 B1 EP 1648607B1 EP 04741090 A EP04741090 A EP 04741090A EP 04741090 A EP04741090 A EP 04741090A EP 1648607 B1 EP1648607 B1 EP 1648607B1
- Authority
- EP
- European Patent Office
- Prior art keywords
- substrate
- stack
- plates
- substrate according
- plate
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Not-in-force
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Classifications
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
- B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0236—Mechanical aspects
- A01N1/0263—Non-refrigerated containers specially adapted for transporting or storing living parts whilst preserving, e.g. cool boxes, blood bags or "straws" for cryopreservation
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0236—Mechanical aspects
- A01N1/0263—Non-refrigerated containers specially adapted for transporting or storing living parts whilst preserving, e.g. cool boxes, blood bags or "straws" for cryopreservation
- A01N1/0268—Carriers for immersion in cryogenic fluid, both for slow-freezing and vitrification, e.g. open or closed "straws" for embryos, oocytes or semen
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/02—Adapting objects or devices to another
- B01L2200/025—Align devices or objects to ensure defined positions relative to each other
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0809—Geometry, shape and general structure rectangular shaped
- B01L2300/0829—Multi-well plates; Microtitration plates
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L9/00—Supporting devices; Holding devices
- B01L9/52—Supports specially adapted for flat sample carriers, e.g. for plates, slides, chips
- B01L9/523—Supports specially adapted for flat sample carriers, e.g. for plates, slides, chips for multisample carriers, e.g. used for microtitration plates
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- F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
- F25—REFRIGERATION OR COOLING; COMBINED HEATING AND REFRIGERATION SYSTEMS; HEAT PUMP SYSTEMS; MANUFACTURE OR STORAGE OF ICE; LIQUEFACTION SOLIDIFICATION OF GASES
- F25D—REFRIGERATORS; COLD ROOMS; ICE-BOXES; COOLING OR FREEZING APPARATUS NOT OTHERWISE PROVIDED FOR
- F25D25/00—Charging, supporting, and discharging the articles to be cooled
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S436/00—Chemistry: analytical and immunological testing
- Y10S436/807—Apparatus included in process claim, e.g. physical support structures
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S436/00—Chemistry: analytical and immunological testing
- Y10S436/807—Apparatus included in process claim, e.g. physical support structures
- Y10S436/809—Multifield plates or multicontainer arrays
Definitions
- the invention relates to a substrate for receiving and storing a plurality of samples, and more particularly to a substrate for the cryopreservation of biological samples.
- the invention also relates to methods for cryopreserving samples having such a substrate.
- biological samples in particular biological tissue, tissue parts, biological cells, cell groups, cell components, cell organelles or biologically relevant macromolecules
- the biological samples are placed in the dissolved or suspended state on a sample substrate which is ready for cryopreservation in a reduced temperature environment, e.g. B. is transferred to a cryogenic tank.
- substrates for cryopreservation are known, which are based on support systems in laboratory technology, such as. B. were developed from microtiter plates.
- Important requirements in the development of the conventional substrates for the cryopreservation consist in the provision of a high absorption capacity, in the adaptation to the freezing and storage conditions and in the flexibility and functionality of the substrates (ability to easily adapt to certain conservation tasks, ability to facilitate sampling in the cryopreserved state).
- a disadvantage of the conventional substrates (sample chambers) for the cryopreservation may be that a compact arrangement z. B. is connected in a cryogenic tank with a likelihood of confusion.
- cryocontainer there may be an unintentional redistribution of substrates in a cryocontainer, which can only be corrected by complex measures for data acquisition.
- cryopreservation storage systems Another requirement for cryopreservation storage systems is that low cost mass production should be possible.
- drawer systems for the orderly storage of substrates in cryogenic containers would be disadvantageous, since they have a complicated structure and their adaptability to specific conservation tasks is limited.
- the object of the invention is to provide an improved substrate for receiving a plurality of samples, with which the disadvantages of conventional substrates (sample chambers) are overcome in particular for cryopreservation and in particular has a compact structure, inexpensive mass-produced and sample storage with reduced likelihood of confusion allows.
- Another object of the invention is to provide improved methods for cryopreserving samples, and more particularly for delivering or withdrawing samples to a substrate, particularly under cryogenic conditions.
- a substrate for receiving a plurality of samples comprising a stack of a plurality of substrate plates.
- the substrate plates are detachably connected as sub-substrates in the stack by at least one anchoring axis.
- the inventive combination of individual substrate plates to a stack has the following advantages. By connecting the substrate plates to the anchoring axis, the order of the substrate plates in the stack is determined. An unintentional rearrangement of the substrate plates is excluded. Furthermore, the substrates serve as mutual cover (closure function). This reliably prevents mutual contamination of different samples. It is also advantageous that the substrate stack with the anchoring axis, which may consist of a single part, can be securely locked against unintentional manipulation.
- the substrate according to the invention has a simplified structure, which can be produced completely from low-temperature materials and suitable for cost-effective, mass production.
- the stack composite comprises at least two substrate plates (or: pallets), of which at least one substrate plate is set up for sample taking.
- a substrate plate for sampling is generally a container or carrier in or on which at least one sample is exposed or covered.
- the geometric shape of the container or carrier may be chosen differently depending on the specific tasks of the substrate.
- a Substrate plate contain one or more cup-shaped or elongated sample chambers.
- the substrate plates are stacked in the stack with a particular stacking direction.
- the anchoring axis preferably runs parallel to the stacking direction. When the substrate plates have a planar shape, accordingly, the stacking direction and the anchoring axis are aligned perpendicular to the substrate plate planes.
- the anchoring axis preferably has a substantially rigid shape as a component; it is preferably also resistant to bending in a state without tensile stress, inherently rigid and dimensionally stable. It is especially for the pivoting of individual plates from the stack preferably exactly (exclusively) provided an anchoring axis.
- each substrate plate has a bearing bore through which the anchoring axis passes.
- the bearing bores of the substrate plates and the anchoring axis form a bearing for the substrate plates, so that advantageously a stable positioning of the substrate plates relative to one another is achieved.
- the bearing bores and the anchoring axis can have any matching, round or angular cross-sectional shape. However, a circular cross-section of the bearing bores is preferred for the positive arrangement of a rotatable anchoring axis.
- the substrate plates have a rectangular shape and the bearing bore is provided in each case in a corner of the substrate plates, the substrate plates are advantageously stacked in relation to at least two plate edges in the stack composite. If further all substrate plates have the same footprint, is advantageously formed a straight, compact substrate plate stack with all-round plate edges.
- a modular construction can be realized with the substrate according to the invention, in which a plurality of substrate plate stacks are in turn stacked and / or row-connected with each other, wherein the composite can in turn be locked by one or more anchoring axes.
- the bearing bore of at least one of the substrate plates in the stacked assembly can have an introduction opening at the edge of the substrate plate, through which opening the bearing bore opens toward the circumference of the substrate plate.
- the provision of the insertion opening means that the bearing bore represents a recess formed in the edge of the respective substrate plate. This allows for lateral attachment or detachment of the substrate plate from the anchoring plate without having to remove all of the substrate plates axially aligned on the anchoring axis.
- the insertion opening of the bearing bore thus increases the flexibility in the application of the substrate according to the invention, in which the respective substrate plates can be freely accessed. It is particularly preferred if all substrate plates are each equipped with the insertion opening on the bearing bore.
- the introduction opening forms at least over a portion of the thickness of the substrate plate, a collar opening having a width which is less than the cross-sectional dimension, in particular less than the diameter of the bearing bore.
- the anchoring axis has at least in sections a thickness such that it can be pushed through the collar opening. It can be provided on the anchoring axis sections of reduced thickness, axially formed according to the position of the collar opening in the stacking direction and / or radially as a cut key surfaces.
- the anchoring axis has a projection at its upper end, a stop for fixing the substrate plates in the stack composite can be formed.
- the projection preferably has a diameter which is greater than the diameter of the bearing bore in the substrate plates.
- the anchoring axis is rotatably arranged in the bearing bores. This advantageously allows, firstly, a suitable alignment of the anchoring axis relative to the collar openings in the stack composite, second, a pivotability of individual substrate plates (see below) and, thirdly, a fixation of the anchoring axis by screwing to a base plate.
- further components may be contained in the stack of the substrate plates which fulfill other functions than the sample holder.
- at least one data storage device, a base plate and / or a cover plate may be provided, each of which preferably has the same outer shape as the substrate plates.
- a data memory can be integrated in the base plate and / or the cover plate, in which information is electronically or optically stored which characterizes the substrate and / or the stored samples.
- the anchoring axis with a lowermost substrate plate or the base plate retractable, z. B. is connected by a screw
- the substrate plate composite can be advantageously clamped between the projection at the upper end of the anchoring axis and corresponding to the lowermost substrate plate or the base plate.
- the state in which all the substrate plates in the stack are mutually fixed is also referred to as the fixing position.
- a displacement of individual substrate plates can be provided perpendicular to the orientation of the anchoring axis, in which case the substrate plate from the composite with the remaining substrate plates and the anchoring axis is solvable.
- the anchoring axis can be transferred by a rotation of the lowered fixing position into a rotational position in which the substrate plates are movable according to a margin in the stacking direction and about the anchoring axis, and / or in a release position, in the at least one Substrate plate can be separated from the stack.
- the stability of the composite of the substrate plates can be increased if according to a further embodiment of the invention engagement means are provided which prevent a lateral displacement of the substrate plates relative to each other, in particular at least in a direction perpendicular to the stacking direction.
- engagement means are provided which prevent a lateral displacement of the substrate plates relative to each other, in particular at least in a direction perpendicular to the stacking direction.
- profiles may be provided on flat side surfaces of the substrate plates, which engage in one another in the substrate composite.
- the profilings consist, for example, of knob-shaped projections on one side of the plate and complementary recesses on the opposite, adjacent side of the plate.
- the mutual engagement of such profiles can be released by a relaxation of the anchoring axis.
- the engagement means comprise a positive sliding guide.
- the sliding guide includes, for example, at least one ridge on a side surface of a substrate plate that cooperates with a groove on a side surface of an adjacent substrate plate.
- other sliding guides such as combinations of cylindrical pins with matching bores or dovetail guides may be provided. With the sliding guides, the substrate plates can be pushed together like drawers and separated from each other.
- the substrate is also referred to as a rotary stack substrate
- the substrate plates are pivotable relative to each other and possibly also displaceable.
- the anchoring axis is a one-piece rod or pin that extends over the entire height of the stack of substrate plates (and possibly provided additional, plate-shaped components) extends.
- the rod has along its length cutting surfaces (key surfaces) that allow insertion or removal of the respective substrate plate for a certain orientation relative to the collar openings of the bearing bores.
- the anchoring axis in this case forms both the stop for a common orientation of the substrate plates and a clamping device for the substrate plate stack.
- the substrate plates are exclusively displaceable relative to each other.
- the anchoring axis preferably includes a plurality of axis segments corresponding to the number of substrate plates (or additional plate-shaped components) in the substrate stack.
- the formation of the anchoring axis from a plurality of axis segments has the following particular advantage. With the number of substrate plates (or additional components in the stack), each equipped with an axis segment, the correct length of the anchoring axis is automatically given.
- Each axle segment has a cylindrical body with a height substantially equal to the thickness of the substrate plates and a diameter corresponding to the diameter of the bearing bores.
- On the upper and lower sides of the axle segments mutually complementary projections and recesses are provided, which engage in the composite stack of substrate plates. Depending on the orientation of z. B. slot-shaped recesses individual substrate plates can be pulled out of the composite of the stack or blocked in the stack.
- the substrate plates each have a compartment arrangement with a multiplicity of cup-shaped sample reservoirs.
- the geometric arrangement of the sample reservoirs can be adapted to the geometric arrangement of micro- or nanotiter plates, as are common in laboratory technology.
- the substrate plates can be equipped in each case with an electronic or optical data memory which is set up to store information about the samples recorded in the respective substrate plate.
- the substrates are completely made of plastic, z. B. TPX, PE, PTFE, PU o. The like.
- the parts of the Sustrate can be inexpensively manufactured by injection molding and then assembled.
- the stack composite can also be miniaturized.
- the substrate plates have side lengths that are less than 10 cm, preferably less than 6 cm.
- the substrate according to the invention can be produced from a plurality of components (in particular anchoring axis, substrate plates) of identical or different plastics which ensure sufficient stability in all operating states and are movable relative to one another , It has surprisingly been found that the plastics used are relatively soft and deformable at room temperature, but nevertheless sufficiently stable. At the low preservation temperatures, however, the plastics are hard and inelastic, while maintaining their relative mobility with adapted thermal expansion coefficients.
- the invention is based on the general technical teaching of depositing samples for cryopreservation in a substrate according to the invention with a stack of plates and freezing them in the stacked composite.
- the formation of the stack can take place before or after the deposition of the samples.
- the loading of the substrate plates after formation of the stack may have the advantage that unintentional permutations of substrate plates are avoided.
- the loading of the substrate plates prior to formation of the stack may have advantages in handling the substrate plates, for example, in a laboratory.
- individual substrate plates in the frozen or thawed state are pivoted out of the stacked composite and / or pushed, so that individual samples can be removed in a targeted manner from the substrate according to the invention.
- an electronic or optical data storage device 50 eg FLASH memory
- the substrate plates 11, 12, 13 are each flat, plate-shaped components with a rectangular basic shape, on the upper side of the compartment assembly 40 with a plurality of sample reservoirs 41, 42, 43 is formed.
- the sample reservoirs 41, 42, 43 are each cup-shaped depressions with a circumferential, circular edge.
- the upper side of the substrate plates (eg, 13 in Figure 2) has a peripheral edge 17 which widens on at least one side to provide the engagement means 30 (see below) and which rises higher above the plane of the plate than the edges of the plates Sample Reservoirs 41, 42, 43.
- a protective film can be stretched over the upper side of the substrate plates and rests on the edge 17.
- the substrate plates 11, 12, 13 are made of plastic or possibly a composite material made of a plastic, in which a metal (for example, aluminum) is embedded.
- a memory such as a magnetic, optical or electronic memory can be integrated (inserted, cast or injected).
- the plates in the stack 10 have manipulation openings 70 (for example 71 in FIG. 2 or 72 in FIG. 9) on at least one side.
- the manipulation openings 70 are used to manipulate manipulation devices, tools or other auxiliary devices with which in particular the transport of the entire substrate or of individual plates is carried out.
- the enlarged partial views of the substrate plates 11, 12 in Figures 4 and 5 show in a corner of the substrate plate in each case the bearing bore 15 which opens via the insertion opening 16 to the periphery of the substrate plates 11, 12 out.
- the insertion opening 16 has a collar opening 18 extending over approximately half the thickness of the substrate plate 11, at which the width of the gap formed by the insertion opening 16 is smaller than the diameter of the bearing bore 15.
- the collar (edge of the collar opening 18) forms a retaining element with the alignment of the anchoring axis relative to the substrate plate (see below).
- a knob-shaped projection 32 is provided in addition to the bearing bore 15 as profiling, which together with a profiling, such as the recess 31 on the adjacent upper side of the adjacent substrate plate 12 ( Figure 5) forms the engaging means 30 of the rotary stack substrate 100.
- the anchoring axis 20 comprises a continuous rod 21 with a specific outer diameter corresponding to the diameter of the bearing bores 15 in the substrate plates and a projection 22 provided at the upper end with a larger diameter.
- the rod 21 cut or key surfaces 23 are provided, on which the thickness of the rod 21 is reduced to the width of the collar opening 18 of the insertion opening 16.
- the key surfaces 23 have an axial length greater than or equal to the length of the collar apertures 18 (in the stacking direction) and an axial distance substantially equal to the spacing of the collar apertures 18 of the substrate plates in the stacking direction.
- a thread 24 is provided at the lower end of the rod 21, a thread 24 is provided.
- the thread 24 can extend over the entire length of the anchoring axis 20, so that it is constructed like a screw. This design allows easy cutting of the desired length of an anchoring axis.
- a slot 25 is provided in the projection 22.
- the anchoring axis 20 is made of plastic or possibly a composite material made of plastic, in which a metal core (for example made of aluminum) is embedded.
- the base plate 60 shown in Figure 7 forms a lowermost support for the stack 10 of the substrate plates 11, 12, 13.
- the base plate 60 has on its upper side a threaded hole 61, which is aligned according to the position of the bearing bores 15 and for receiving the thread 24 of Anchoring axis 20 is set up. Furthermore, it is analogous to Recess 31 is provided a recess 62 on the upper side of the base plate 60.
- the base plate 60 has a recess 63 for receiving a magnetic, electronic or optical data memory (not shown).
- the data memory is inserted into the recess 63 and fixed by projections 64 at the edges of the recess 63. Alternatively, pouring or injecting the data memory may be provided.
- the recess 63 has at least one lateral opening, through which on the one hand, even with a composite stack, the data memory can be inserted and, on the other hand, an electrical connection for the data memory can be carried out.
- the reference numeral 65 refers to an interface opening (see also FIG. 12).
- the data memory is formed, for example, by a Compact FLASH memory
- a plug with contact pins for connection to the Compact FLASH memory can be inserted through the interface opening 65 and optionally at least temporarily fixed laterally to the base plate (eg with a clip Connection). Via the interface, the data memory can be connected to an external control device.
- the attachment of the data memory described on the example of the base plate 60 may also be provided on at least one of the substrate plates or the cover plate.
- FIG. 8 illustrates the first step in constructing a rotary stack substrate 100 according to the invention.
- the anchoring axis 20 is loosely screwed into the base part 60, so that the key surfaces 23 are perpendicular to the introduction opening 16.
- the release position the key surfaces 23 are along the length of the anchoring axis each at a height above the base plate, that the collar openings (18) of the substrate plates in the stack are each aligned with the key surfaces 23.
- the anchoring axis can be inserted through the insertion openings 16 in the bearing bore or pushed out of this.
- the lowermost substrate plate 11 which may already be loaded with samples, is pushed onto the base plate 60. Since the lowermost key surface 23 is aligned properly, the substrate plate 11 can be advanced until the anchoring axis 20 passes through the bearing bore 15. Subsequently, further substrate plates are pushed according to the length of the anchoring axis 20 used.
- the anchoring axis 20 is initially after completion of the stack 10 in the raised state of the release position.
- the anchoring axis z. B. With a screwdriver, which engages in the slot 25 of the projection 22 (see Figure 6), the anchoring axis 20 is lowered.
- the alignment of the key surfaces 23 with the collar openings 18 along the length of the anchoring axis 20 is lost.
- the substrate plates 11, 12, 13 can no longer be separated from the stack 10.
- This condition is also referred to as the rotational position of the anchoring axis 20.
- the clearance of the substrate plates 11, 12, 13 is greater than the height of the profilings 31, 32, so that the substrate plates 11, 12, 13 can be pivoted out of the stack about the anchoring axis 20.
- the anchoring axis 20 is firmly screwed into the base plate 60. This condition is also referred to as the fixing position of the anchoring axis 20. In the fixing position, the substrate plates are pressed together, so that the engagement means 31, 32 interlock and block further displacement or pivoting of the substrate plates.
- the freezing and storage of the substrate 100 may be carried out, for example, at the temperature of the liquid nitrogen or in the vapor of the liquid nitrogen (normal pressure).
- the anchoring axis 20 can be converted by loosening the screw on the base plate 60 in the rotational position.
- the engagement means 31, 32 are released, so that the substrate plate 12 is laterally pivotable about the anchoring axis 20 to the outside (Fig. 1).
- the rotation of the anchoring axis 20 and / or the substrate plate 12 may be selected so that the key surfaces 23 cooperate with the insertion openings 16, so that the substrate plate 12 can be separated from the stack 10.
- 1 to 20 substrate plates are arranged one above the other. Depending on the desired number of plates an anchoring axis 20 is used with a suitable length.
- the anchoring axis between the release, rotational and fixing positions solely by a rotation can be adjusted by screwing into the base plate.
- the increase in the screw is determined by the pitch of the thread 24. This can advantageously be determined by the number of revolutions of the anchoring axis the transfer between the different positions.
- At least one information carrier is preferably provided on a substrate stack, which is provided by the above-mentioned o.
- Data storage and / or by additional storage media, such as a bar code is formed.
- FIG. 9-12 A preferred embodiment of a sliding stack substrate 200 is illustrated in Figures 9-12.
- the engaging means 30 are formed as a positive sliding guide, for example, of ridges 33 and grooves 34, which are formed as straight guide rails complementary to each other at the edges of the upper and lower sides of the substrate plates.
- ridges 33 and grooves 34 are formed as straight guide rails complementary to each other at the edges of the upper and lower sides of the substrate plates.
- a one-piece anchoring axis 20 may be provided with the key surfaces 23 to prevent completion of the stack 10, a further displacement of the substrate plates.
- an anchoring axis 20 formed of axle segments 26 is used in the sliding stack substrate. This has the advantage that the length of the anchoring axis 20 is easily adjusted by the number of axis segments 26 used to match the number of substrate plates.
- Each axle segment 26 comprises a cylindrical body, on whose upper and lower sides complementary to one another, slit-shaped recesses 27 and projections 28 are formed.
- a projection 28 engages each recess 27 of the underlying axle segment 26.
- the axle segments 26 are rotatably arranged in the bearing bores 15 (see FIG. 11).
- the substrate plates are prefabricated with the axis segments.
- the axis segments are pressed into the bearing bores of the substrate plates at room temperature.
- the axle segments 26 can hardly be removed from the bearing bores 15 without destruction.
- a base part 60 (FIG. 12) is provided, in which an axis segment 26 is correspondingly arranged.
- the substrate plates and anchoring axes of the substrates 100, 200 are preferably produced by injection molding from TPX, PE, PTFE or the like.
- the side lengths of the substrate plates are, for example, in the range of 10 mm to 20 cm or above, such as. B. 50 cm or 80 cm.
- the thickness of the substrate plates is, for example, 4 mm to 5 cm or more.
- the number of sample reservoirs 41, 42, 43 per substrate plate is dependent on the size of the substrate plate and the sample reservoirs and is, for example, 20 to 200 for smaller formats. For larger formats For example, the number can be significantly higher, ranging from 5,000 to 10,000.
- the size and shape of the sample reservoirs depends on the biological samples (in particular biological tissue, tissue parts, biological cells, cell groups, cell components, cell organelles or biologically relevant macromolecules) which are to be stored.
- biological samples in particular biological tissue, tissue parts, biological cells, cell groups, cell components, cell organelles or biologically relevant macromolecules
- substrate plates may be combined with different base areas in the stack.
- the base in the stack can be smaller towards the top.
- the anchoring axes and bearing bores each have a round cross-section. It can also be provided an anchoring axis with a polygonal cross-section.
- the key surfaces can be aligned differently relative to one another, so that in each case one substrate plate is released during the rotation of the anchoring axis and the others are blocked.
Abstract
Description
Die Erfindung betrifft ein Substrat zur Aufnahme und Lagerung einer Vielzahl von Proben und insbesondere ein Substrat für die Kryokonservierung biologischer Proben. Die Erfindung betrifft auch Verfahren zur Kryokonservierung von Proben mit einem derartigen Substrat.The invention relates to a substrate for receiving and storing a plurality of samples, and more particularly to a substrate for the cryopreservation of biological samples. The invention also relates to methods for cryopreserving samples having such a substrate.
Es ist bekannt, biologische Proben (insbesondere biologisches Gewebe, Gewebeteile, biologische Zellen, Zellgruppen, Zellbestandteile, Zellorganellen oder biologisch relevante Makromoleküle) dauerhaft im gefrorenen Zustand zu lagern (Kryokonservierung). Die biologischen Proben werden im gelösten oder suspendierten Zustand auf einem Probensubstrat angeordnet, das zur Kryokonservierung in eine Umgebung reduzierter Temperatur, z. B. in einen Kryotank überführt wird.It is known to permanently store biological samples (in particular biological tissue, tissue parts, biological cells, cell groups, cell components, cell organelles or biologically relevant macromolecules) in a frozen state (cryopreservation). The biological samples are placed in the dissolved or suspended state on a sample substrate which is ready for cryopreservation in a reduced temperature environment, e.g. B. is transferred to a cryogenic tank.
Aus der Praxis sind verschiedene Formen von Substraten für die Kryokonservierung bekannt, die auf der Grundlage von Trägersystemen in der Labortechnik, wie z. B. aus Mikrotiterplatten entwickelt wurden. Wichtige Anforderungen bei der Entwicklung der herkömmlichen Substrate für die Kryokonservierung bestanden in der Bereitstellung einer hohen Aufnahmekapazität, in der Anpassung an die Einfrier- und Lagerungsbedingungen und in der Flexibilität und Funktionalität der Substrate (Fähigkeit zur einfachen Anpassung an bestimmte Konservierungsaufgaben, Fähigkeit zur erleichterten Probenentnahme im kryokonservierten Zustand). Nachteilig an den herkömmlichen Substraten (Probenkammern) für die Kryokonservierung kann jedoch sein, dass eine kompakte Anordnung z. B. in einem Kryotank mit einer Verwechslungsgefahr verbunden ist.From practice, various forms of substrates for cryopreservation are known, which are based on support systems in laboratory technology, such as. B. were developed from microtiter plates. Important requirements in the development of the conventional substrates for the cryopreservation consist in the provision of a high absorption capacity, in the adaptation to the freezing and storage conditions and in the flexibility and functionality of the substrates (ability to easily adapt to certain conservation tasks, ability to facilitate sampling in the cryopreserved state). However, a disadvantage of the conventional substrates (sample chambers) for the cryopreservation may be that a compact arrangement z. B. is connected in a cryogenic tank with a likelihood of confusion.
Es kann bspw. zu einer unbeabsichtigten Umverteilung von Substraten in einem Kryobehälter kommen, die nur durch aufwändige Maßnahmen zur Datenerfassung korrigiert werden kann.For example, there may be an unintentional redistribution of substrates in a cryocontainer, which can only be corrected by complex measures for data acquisition.
Eine weitere Anforderung an Aufbewahrungssysteme für die Kryokonservierung besteht darin, dass eine kostengünstige Massenproduktion möglich sein soll. Mit Blick auf dieses Kriterium wären bspw. Schubladensysteme zur geordneten Ablage von Substraten in Kryobehältern nachteilig, da sie einen komplizierten Aufbau besitzen und ihre Anpassungsfähigkeit an konkrete Konservierungsaufgaben beschränkt ist.Another requirement for cryopreservation storage systems is that low cost mass production should be possible. With regard to this criterion, for example, drawer systems for the orderly storage of substrates in cryogenic containers would be disadvantageous, since they have a complicated structure and their adaptability to specific conservation tasks is limited.
Die genannten Probleme treten nicht nur bei Substraten zur Probenhalterung für die Kryokonservierung, sondern allgemein bei Probenträgern für flüssige (suspendierte oder gelöste) oder partikelförmige Proben biologischen oder synthetischen Ursprungs für Bearbeitungs-, Reaktions- oder Lagerungszwecke auf.The problems mentioned occur not only with substrates for the sample holder for cryopreservation, but generally with sample carriers for liquid (suspended or dissolved) or particulate samples of biological or synthetic origin for processing, reaction or storage purposes.
Die Aufgabe der Erfindung ist es, ein verbessertes Substrat zur Aufnahme einer Vielzahl von Proben bereitzustellen, mit dem die Nachteile der herkömmlichen Substrate (Probenkammern) insbesondere zur Kryokonservierung überwunden werden und das insbesondere einen kompakten Aufbau besitzt, kostengünstig als Massenprodukt herstellbar ist und eine Probenablage mit verminderter Verwechslungsgefahr ermöglicht. Eine weitere Aufgabe der Erfindung ist die Bereitstellung verbesserter Verfahren zur Kryokonservierung von Proben und insbesondere zur Zuführung oder Entnahme von Proben an einem Substrat insbesondere unter Tieftemperaturbedingungen.The object of the invention is to provide an improved substrate for receiving a plurality of samples, with which the disadvantages of conventional substrates (sample chambers) are overcome in particular for cryopreservation and in particular has a compact structure, inexpensive mass-produced and sample storage with reduced likelihood of confusion allows. Another object of the invention is to provide improved methods for cryopreserving samples, and more particularly for delivering or withdrawing samples to a substrate, particularly under cryogenic conditions.
Diese Aufgaben werden durch Substrate und Verfahren mit den Merkmalen gemäß den Patentansprüchen 1 oder 25 gelöst.These objects are achieved by substrates and methods having the features according to
Vorteilhafte Ausführungsformen und Anwendungen der Erfindung ergeben sich aus den abhängigen Ansprüchen.Advantageous embodiments and applications of the invention will become apparent from the dependent claims.
Vorrichtungsbezogen wird die o. g. Aufgabe durch die allgemeine technische Lehre gelöst, ein Substrat zur Aufnahme einer Vielzahl von Proben bereitzustellen, das einen Stapelverbund aus einer Vielzahl von Substratplatten umfasst. Die Substratplatten sind als Teilsubstrate im Stapel durch mindestens eine Verankerungsachse lösbar verbunden. Die erfindungsgemäße Kombination einzelner Substratplatten zu einem Stapel besitzt die folgenden Vorteile. Durch die Verbindung der Substratplatten mit der Verankerungsachse ist die Reihenfolge der Substratplatten im Stapel festgelegt. Eine unbeabsichtigte Umordnung der Substratplatten ist ausgeschlossen. Des Weiteren dienen die Substrate als gegenseitige Abdeckung (Verschlussfunktion). Dadurch wird eine gegenseitige Kontamination verschiedener Proben sicher verhindert. Vorteilhaft ist ferner, dass der Substratstapel mit der Verankerungsachse, die aus einem einzigen Teil bestehen kann, sicher gegen unbeabsichtigte Manipulationen verriegelt werden kann. Das erfindungsgemäße Substrat besitzt einen vereinfachten Aufbau, der vollständig aus tieftemperaturtauglichen Materialien herstellbar und für eine kostengünstige, massenhafte Produktion geeignet ist.Device related is the o. G. Problem solved by the general technical teaching to provide a substrate for receiving a plurality of samples comprising a stack of a plurality of substrate plates. The substrate plates are detachably connected as sub-substrates in the stack by at least one anchoring axis. The inventive combination of individual substrate plates to a stack has the following advantages. By connecting the substrate plates to the anchoring axis, the order of the substrate plates in the stack is determined. An unintentional rearrangement of the substrate plates is excluded. Furthermore, the substrates serve as mutual cover (closure function). This reliably prevents mutual contamination of different samples. It is also advantageous that the substrate stack with the anchoring axis, which may consist of a single part, can be securely locked against unintentional manipulation. The substrate according to the invention has a simplified structure, which can be produced completely from low-temperature materials and suitable for cost-effective, mass production.
Der Stapelverbund umfasst mindestens zwei Substratplatten (oder: Paletten), von denen wenigstens eine Substratplatte zur Probenaufnahme eingerichtet ist. Eine Substratplatte zur Probenaufnahme ist allgemein ein Behältnis oder Träger, in oder auf dem wenigstens eine Probe freiliegend oder abgedeckt angeordnet ist. Die geometrische Form des Behältnisses oder Trägers kann je nach den konkreten Aufgaben des Substrats verschieden gewählt sein. Beispielsweise kann eine Substratplatte eine oder mehrere topfförmige oder langgestreckte Probenkammern enthalten.The stack composite comprises at least two substrate plates (or: pallets), of which at least one substrate plate is set up for sample taking. A substrate plate for sampling is generally a container or carrier in or on which at least one sample is exposed or covered. The geometric shape of the container or carrier may be chosen differently depending on the specific tasks of the substrate. For example, a Substrate plate contain one or more cup-shaped or elongated sample chambers.
Die Substratplatten sind im Stapel mit einer bestimmten Stapelrichtung übereinander angeordnet. Die Verankerungsachse verläuft vorzugsweise parallel zu der Stapelrichtung. Wenn die Substratplatten eine ebene Form besitzen, sind entsprechend die Stapelrichtung und die Verankerungsachse senkrecht zu den Substratplattenebenen ausgerichtet. Die Verankerungsachse weist als Bauteil vorzugsweise eine im Wesentlichen starre Gestalt auf, sie ist vorzugsweise auch in einem Zustand ohne Zugspannung biegefest, eigensteif und formstabil. Es ist insbesondere für das Schwenken einzelner Platten aus dem Stapel vorzugsweise genau (ausschließlich) eine Verankerungsachse vorgesehen.The substrate plates are stacked in the stack with a particular stacking direction. The anchoring axis preferably runs parallel to the stacking direction. When the substrate plates have a planar shape, accordingly, the stacking direction and the anchoring axis are aligned perpendicular to the substrate plate planes. The anchoring axis preferably has a substantially rigid shape as a component; it is preferably also resistant to bending in a state without tensile stress, inherently rigid and dimensionally stable. It is especially for the pivoting of individual plates from the stack preferably exactly (exclusively) provided an anchoring axis.
Gemäß einer bevorzugten Ausführungsform der Erfindung besitzt jede Substratplatte eine Lagerbohrung, durch die die Verankerungsachse hindurchtritt. Die Lagerbohrungen der Substratplatten und die Verankerungsachse bilden eine Lagerung für die Substratplatten, so dass vorteilhafterweise eine stabile Positionierung der Substratplatten relativ zueinander erzielt wird. Die Lagerbohrungen und die Verankerungsachse können jede passende, runde oder eckige Querschnittsform besitzen.
Ein kreisrunder Querschnitt der Lagerbohrungen wird jedoch für die formschlüssige Anordnung einer drehbaren Verankerungsachse bevorzugt.According to a preferred embodiment of the invention, each substrate plate has a bearing bore through which the anchoring axis passes. The bearing bores of the substrate plates and the anchoring axis form a bearing for the substrate plates, so that advantageously a stable positioning of the substrate plates relative to one another is achieved. The bearing bores and the anchoring axis can have any matching, round or angular cross-sectional shape.
However, a circular cross-section of the bearing bores is preferred for the positive arrangement of a rotatable anchoring axis.
Wenn gemäß einer weiteren bevorzugten Ausführungsform die Substratplatten eine rechteckige Form besitzen und die Lagerbohrung jeweils in einer Ecke der Substratplatten vorgesehen ist, werden die Substratplatten vorteilhafterweise im Stapelverbund in Bezug auf mindestens zwei Plattenränder fluchtend übereinander angeordnet. Wenn des Weiteren alle Substratplatten die gleiche Grundfläche besitzen, wird vorteilhafterweise ein gerader, kompakter Substratplattenstapel mit allseits ausgerichteten Plattenrändern gebildet.If according to a further preferred embodiment, the substrate plates have a rectangular shape and the bearing bore is provided in each case in a corner of the substrate plates, the substrate plates are advantageously stacked in relation to at least two plate edges in the stack composite. If further all substrate plates have the same footprint, is advantageously formed a straight, compact substrate plate stack with all-round plate edges.
Vorteilhafterweise kann mit dem erfindungsgemäßen Substrat eine Modulkonstruktion realisiert werden, in der eine Vielzahl von Substratplattenstapeln wiederum stapel- und/oder reihenweise miteinander verbunden werden, wobei der Verbund wiederum durch eine oder mehrere Verankerungsachsen verriegelt werden kann.Advantageously, a modular construction can be realized with the substrate according to the invention, in which a plurality of substrate plate stacks are in turn stacked and / or row-connected with each other, wherein the composite can in turn be locked by one or more anchoring axes.
Gemäß einer besonders vorteilhaften Variante der Erfindung kann vorgesehen sein, dass die Lagerbohrung mindestens einer der Substratplatten im Stapelverbund am Rand der Substratplatte eine Einführungsöffnung aufweist, durch die sich die Lagerbohrung zum Umfang der Substratplatte hin öffnet. Die Bereitstellung der Einführungsöffnung bedeutet, dass die Lagerbohrung eine im Rand der jeweiligen Substratplatte geformte Ausnehmung darstellt. Dies ermöglicht ein laterales Ansetzen oder Abnehmen der Substratplatte von der Verankerungsplatte, ohne dass die axial auf der Verankerungsachse aufgereihten Substratplatten sämtlich abgenommen werden müssen. Die Einführungsöffnung der Lagerbohrung erhöht damit die Flexibilität bei der Anwendung des erfindungsgemäßen Substrats, in dem auf die jeweiligen Substratplatten frei zugegriffen werden kann. Besonders bevorzugt ist es, wenn alle Substratplatten jeweils mit der Einführungsöffnung an der Lagerbohrung ausgestattet sind.According to a particularly advantageous variant of the invention, provision can be made for the bearing bore of at least one of the substrate plates in the stacked assembly to have an introduction opening at the edge of the substrate plate, through which opening the bearing bore opens toward the circumference of the substrate plate. The provision of the insertion opening means that the bearing bore represents a recess formed in the edge of the respective substrate plate. This allows for lateral attachment or detachment of the substrate plate from the anchoring plate without having to remove all of the substrate plates axially aligned on the anchoring axis. The insertion opening of the bearing bore thus increases the flexibility in the application of the substrate according to the invention, in which the respective substrate plates can be freely accessed. It is particularly preferred if all substrate plates are each equipped with the insertion opening on the bearing bore.
Weitere Vorteile können sich ergeben, wenn die Einführungsöffnung ein Einsetzen oder Abnehmen der jeweiligen Substratplatte nur bei einer vorgegebenen geometrischen Ausrichtung der Substratplatte relativ zur Verankerungsachse ermöglicht. Hierzu sind die folgenden Maßnahmen vorgesehen. Die Einführungsöffnung bildet wenigstens über einen Teil der Dicke der Substratplatte eine Kragenöffnung mit einer Breite, die geringer als die Querschnittsdimension, insbesondere geringer als der Durchmesser der Lagerbohrung ist. Die Verankerungsachse besitzt mindestens in Teilabschnitten eine Dicke derart, dass sie durch die Kragenöffnung geschoben werden kann. Es können an der Verankerungsachse Teilabschnitte verminderter Dicke vorgesehen sein, axial entsprechend der Position der Kragenöffnung in Stapelrichtung und/oder radial als eingeschnittene Schlüsselflächen gebildet sind. Wenn die Substratplatte und die Verankerungsachse relativ zueinander so ausgerichtet werden, dass die Kragenöffnung und der Teilabschnitt mit verminderter Dicke zueinander ausgerichtet sind, kann die Substratplatte in einer Richtung senkrecht zur Verankerungsachse von dieser abgezogen werden.Further advantages may be obtained if the insertion opening permits insertion or removal of the respective substrate plate only with a predetermined geometric orientation of the substrate plate relative to the anchoring axis. For this purpose, the following measures are provided. The introduction opening forms at least over a portion of the thickness of the substrate plate, a collar opening having a width which is less than the cross-sectional dimension, in particular less than the diameter of the bearing bore. The anchoring axis has at least in sections a thickness such that it can be pushed through the collar opening. It can be provided on the anchoring axis sections of reduced thickness, axially formed according to the position of the collar opening in the stacking direction and / or radially as a cut key surfaces. When the substrate plate and the anchoring axis are aligned relative to each other so that the collar opening and the reduced thickness portion are aligned with each other, the substrate plate can be withdrawn therefrom in a direction perpendicular to the anchoring axis.
Wenn gemäß einer weiteren Ausführungsform der Erfindung die Verankerungsachse an ihrem oberen Ende eine Auskragung aufweist, kann ein Anschlag zur Fixierung der Substratplatten im Stapelverbund gebildet werden. Die Auskragung besitzt vorzugsweise einen Durchmesser, der größer als der Durchmesser der Lagerbohrung in den Substratplatten ist.If according to a further embodiment of the invention, the anchoring axis has a projection at its upper end, a stop for fixing the substrate plates in the stack composite can be formed. The projection preferably has a diameter which is greater than the diameter of the bearing bore in the substrate plates.
Gemäß eine weiteren Variante ist die Verankerungsachse in den Lagerbohrungen drehbar angeordnet. Dies ermöglicht vorteilhafterweise erstens eine passende Ausrichtung der Verankerungsachse relativ zu den Kragenöffnungen im Stapelverbund, zweitens eine Verschwenkbarkeit einzelner Substratplatten (siehe unten) und drittens eine Fixierung der Verankerungsachse durch Festschrauben an einer Basisplatte.According to a further variant, the anchoring axis is rotatably arranged in the bearing bores. This advantageously allows, firstly, a suitable alignment of the anchoring axis relative to the collar openings in the stack composite, second, a pivotability of individual substrate plates (see below) and, thirdly, a fixation of the anchoring axis by screwing to a base plate.
Gemäß einer weiteren vorteilhaften Abwandlung der Erfindung können im Stapel der Substratplatten weitere Komponenten enthalten sein, die andere Funktionen als die Probenaufnahme erfüllen. Beispielsweise kann mindestens eine Datenspeichereinrichtung, eine Basisplatte und/oder eine Abdeckplatte vorgesehen sein, die jeweils vorzugsweise die gleiche äußere Form wie die Substratplatten besitzen. Vorteilhafterweise kann in der Basisplatte und/oder der Abdeckplatte ein Datenspeicher integriert sein, in dem elektronisch oder optisch Informationen gespeichert werden, die das Substrat und/oder die gespeicherten Proben charakterisieren.According to a further advantageous modification of the invention, further components may be contained in the stack of the substrate plates which fulfill other functions than the sample holder. For example, at least one data storage device, a base plate and / or a cover plate may be provided, each of which preferably has the same outer shape as the substrate plates. Advantageously, a data memory can be integrated in the base plate and / or the cover plate, in which information is electronically or optically stored which characterizes the substrate and / or the stored samples.
Wenn gemäß einer bevorzugten Ausführungsform der Erfindung die Verankerungsachse mit einer untersten Substratplatte oder der Basisplatte versenkbar, z. B. durch eine Schraubverbindung verbunden ist, kann der Substratplattenverbund vorteilhafterweise zwischen der Auskragung am oberen Ende der Verankerungsachse und entsprechend der untersten Substratplatte oder der Basisplatte eingeklemmt werden. Der Zustand, in dem alle Substratplatten im Stapel gegenseitig fixiert sind, wird auch als Fixierposition bezeichnet.If, according to a preferred embodiment of the invention, the anchoring axis with a lowermost substrate plate or the base plate retractable, z. B. is connected by a screw, the substrate plate composite can be advantageously clamped between the projection at the upper end of the anchoring axis and corresponding to the lowermost substrate plate or the base plate. The state in which all the substrate plates in the stack are mutually fixed is also referred to as the fixing position.
Besondere Vorteile für den Zugriff auf einzelne Substratplatten oder einzelne Proben auf den Substratplatten ergeben sich, wenn die Substratplatten aus dem Stapel um die Verankerungsachse verschwenkbar sind. Alternativ oder zusätzlich kann auch eine Verschiebbarkeit einzelner Substratplatten senkrecht zur Ausrichtung der Verankerungsachse vorgesehen sein, wobei in diesem Fall die Substratplatte aus dem Verbund mit den übrigen Substratplatten und der Verankerungsachse lösbar ist. Hierzu ist vorzugsweise vorgesehen, dass die Verankerungsachse durch eine Verdrehung von der abgesenkten Fixierposition in eine Drehposition, in der die Substratplatten entsprechend einem Spielraum in Stapelrichtung beweglich und um die Verankerungsachse verschwenkbar sind, und/oder in eine Freigabeposition überführt werden kann, in der mindestens eine Substratplatte vom Stapel getrennt werden kann.Particular advantages for access to individual substrate plates or individual samples on the substrate plates arise when the substrate plates are pivotable out of the stack about the anchoring axis. Alternatively or additionally, a displacement of individual substrate plates can be provided perpendicular to the orientation of the anchoring axis, in which case the substrate plate from the composite with the remaining substrate plates and the anchoring axis is solvable. For this purpose, it is preferably provided that the anchoring axis can be transferred by a rotation of the lowered fixing position into a rotational position in which the substrate plates are movable according to a margin in the stacking direction and about the anchoring axis, and / or in a release position, in the at least one Substrate plate can be separated from the stack.
Die Stabilität des Verbundes der Substratplatten kann erhöht werden, wenn gemäß einer weiteren Ausführungsform der Erfindung Eingriffsmittel vorgesehen sind, die eine laterale Verschiebung der Substratplatten relativ zueinander, insbesondere mindestens in einer Richtung senkrecht zur Stapelrichtung verhindern. Beispielsweise können auf ebenen Seitenflächen der Substratplatten Profilierungen vorgesehen sein, die im Substratverbund ineinander greifen. Die Profilierungen bestehen bspw. aus noppenförmigen Vorsprüngen auf einer Plattenseite und komplementären Ausnehmungen auf der entgegengesetzten, angrenzenden Plattenseite. Vorteilhafterweise kann der gegenseitige Eingriff derartiger Profilierungen durch eine Lockerung der Verankerungsachse freigegeben werden.The stability of the composite of the substrate plates can be increased if according to a further embodiment of the invention engagement means are provided which prevent a lateral displacement of the substrate plates relative to each other, in particular at least in a direction perpendicular to the stacking direction. For example, profiles may be provided on flat side surfaces of the substrate plates, which engage in one another in the substrate composite. The profilings consist, for example, of knob-shaped projections on one side of the plate and complementary recesses on the opposite, adjacent side of the plate. Advantageously, the mutual engagement of such profiles can be released by a relaxation of the anchoring axis.
Gemäß einer abgewandelten Variante umfassen die Eingriffsmittel eine formschlüssige Schiebeführung. Die Schiebeführung umfasst zum Beispiel mindestens einen Steg auf einer Seitenfläche einer Substratplatte, der mit einer Nut auf einer Seitenfläche einer angrenzenden Substratplatte zusammenwirkt. Anstelle der ineinander greifenden Stege und Nuten an den Rändern der im Stapel benachbarten Seitenflächen können andere Schiebeführungen, wie zum Beispiel Kombinationen aus Zylinderstiften mit passenden Bohrungen oder Schwalbenschwanzführungen vorgesehen sein. Mit den Schiebeführungen können die Substratplatten wie Schubladen zusammengeschoben und voneinander getrennt werden.According to a modified variant, the engagement means comprise a positive sliding guide. The sliding guide includes, for example, at least one ridge on a side surface of a substrate plate that cooperates with a groove on a side surface of an adjacent substrate plate. Instead of the interlocking webs and grooves on the edges of the side surfaces adjacent in the stack, other sliding guides, such as combinations of cylindrical pins with matching bores or dovetail guides may be provided. With the sliding guides, the substrate plates can be pushed together like drawers and separated from each other.
Bei der Realisierung der Erfindung werden zwei grundsätzliche Ausführungsformen unterschieden. Im ersten Fall, in dem das Substrat auch als Drehstapel-Substrat bezeichnet ist, sind die Substratplatten relativ zueinander verschwenkbar und ggf. auch verschiebbar. Beim Drehstapel-Substrat ist die Verankerungsachse ein einstückiger Stab oder Stift, der sich über die gesamte Höhe des Stapels aus Substratplatten (und aus ggf. vorgesehenen zusätzlichen, plattenförmigen Komponenten) erstreckt. Vorteilhafterweise besitzt der Stab entlang seiner Länge Schnittflächen (Schlüsselflächen), die für eine bestimmte Ausrichtung relativ zu den Kragenöffnungen der Lagerbohrungen ein Einsetzen oder Abziehen der jeweiligen Substratplatte ermöglichen. Vorteilhafterweise bildet die Verankerungsachse in diesem Fall sowohl den Anschlag für eine gemeinsame Ausrichtung der Substratplatten als auch eine Spanneinrichtung für den Substratplattenstapel.In the realization of the invention, two basic embodiments are distinguished. In the first case, in which the substrate is also referred to as a rotary stack substrate, the substrate plates are pivotable relative to each other and possibly also displaceable. In the rotary stack substrate, the anchoring axis is a one-piece rod or pin that extends over the entire height of the stack of substrate plates (and possibly provided additional, plate-shaped components) extends. Advantageously, the rod has along its length cutting surfaces (key surfaces) that allow insertion or removal of the respective substrate plate for a certain orientation relative to the collar openings of the bearing bores. Advantageously, the anchoring axis in this case forms both the stop for a common orientation of the substrate plates and a clamping device for the substrate plate stack.
Im zweiten Fall, in dem das Substrat auch als Schiebestapel-Substrat bezeichnet wird, sind die Substratplatten relativ zueinander ausschließlich verschiebbar. Beim Schiebestapel-Substrat umfasst die Verankerungsachse vorzugsweise eine Vielzahl von Achsensegmenten entsprechend der Anzahl von Substratplatten (oder zusätzlichen, plattenförmigen Komponenten) im Substratstapel. Die Bildung der Verankerungsachse aus einer Vielzahl von Achsensegmenten besitzt den folgenden besonderen Vorteil. Mit der Zahl der Substratplatten (oder zusätzlichen Komponenten im Stapel), die jeweils mit einem Achsensegment ausgestattet sind, ist automatisch die richtige Länge der Verankerungsachse vorgegeben.In the second case, in which the substrate is also referred to as a sliding stack substrate, the substrate plates are exclusively displaceable relative to each other. In the sliding stack substrate, the anchoring axis preferably includes a plurality of axis segments corresponding to the number of substrate plates (or additional plate-shaped components) in the substrate stack. The formation of the anchoring axis from a plurality of axis segments has the following particular advantage. With the number of substrate plates (or additional components in the stack), each equipped with an axis segment, the correct length of the anchoring axis is automatically given.
Jedes Achsensegment besitzt einen zylinderförmigen Körper mit einer Höhe, die im wesentlichen der Dicke der Substratplatten entspricht und einem Durchmesser, der dem Durchmesser der Lagerbohrungen entspricht. An den Ober- und Unterseiten der Achsensegmente sind zueinander komplementäre Vorsprünge und Ausnehmungen vorgesehen, die im zusammengesetzten Stapel von Substratplatten ineinander greifen. Je nach der Ausrichtung der z. B. schlitzförmigen Ausnehmungen können einzelne Substratplatten aus dem Verbund des Stapels gezogen oder im Stapel blockiert werden.Each axle segment has a cylindrical body with a height substantially equal to the thickness of the substrate plates and a diameter corresponding to the diameter of the bearing bores. On the upper and lower sides of the axle segments mutually complementary projections and recesses are provided, which engage in the composite stack of substrate plates. Depending on the orientation of z. B. slot-shaped recesses individual substrate plates can be pulled out of the composite of the stack or blocked in the stack.
Weitere Vorteile der Erfindung in Bezug auf die Probenhandhabung können sich ergeben, wenn die Substratplatten jeweils eine Kompartimentanordnung mit einer Vielzahl von topfförmigen Probenreservoiren aufweisen. Die geometrische Anordnung der Probenreservoire kann an die geometrische Anordnung an Mikro- oder Nanotiterplatten angepasst werden, wie sie in der Labortechnik üblich sind. Des Weiteren können die Substratplatten an sich jeweils mit einem elektronischen oder optischen Datenspeicher ausgestattet sein, der zur Speicherung von Informationen über die in der jeweiligen Substratplatte aufgenommenen Proben eingerichtet ist.Further advantages of the invention with regard to sample handling may result if the substrate plates each have a compartment arrangement with a multiplicity of cup-shaped sample reservoirs. The geometric arrangement of the sample reservoirs can be adapted to the geometric arrangement of micro- or nanotiter plates, as are common in laboratory technology. Furthermore, the substrate plates can be equipped in each case with an electronic or optical data memory which is set up to store information about the samples recorded in the respective substrate plate.
Besondere Vorteile für die Anwendung der Erfindung der Kryokonservierung ergeben sich, wenn die Substrate vollständig aus Kunststoff, z. B. TPX, PE, PTFE, PU o. dgl. bestehen. In diesem Fall können die Teile der Sustrate kostengünstig mit einem Spritzgussverfahren hergestellt und anschließend zusammengesetzt werden. Vorteilhafterweise kann der Stapelverbund auch miniaturisiert werden. Beispielsweise besitzen die Substratplatten Seitenlängen, die geringer als 10 cm, vorzugsweise kleiner als 6 cm sind.Particular advantages for the application of the invention of the cryopreservation arise when the substrates are completely made of plastic, z. B. TPX, PE, PTFE, PU o. The like. Exist. In this case, the parts of the Sustrate can be inexpensively manufactured by injection molding and then assembled. Advantageously, the stack composite can also be miniaturized. For example, the substrate plates have side lengths that are less than 10 cm, preferably less than 6 cm.
Ein wichtiger und bei Substraten für die Kryokonservierung bisher unerreichter Vorteil der Erfindung besteht darin, dass das erfindungsgemäße Substrat aus mehreren Komponenten (insbesondere Verankerungsachse, Substratplatten) aus gleichen oder verschiedenen Kunststoffen herstellbar ist, die in allen Betriebszuständen eine ausreichende Stabilität gewährleisten und relativ zueinander beweglich sind. Es hat sich überraschenderweise herausgestellt, dass die verwendeten Kunststoffe bei Raumtemperatur zwar relativ weich und verformbar, aber dennoch ausreichend stabil sind. Bei den tiefen Konservierungstemperaturen hingegen sind die Kunststoffe hart und unelastisch, wobei sie bei angepassten thermischen Ausdehnungskoeffizienten ihre relative Beweglichkeit bewahren.An important advantage of the invention which has hitherto been unrivaled in substrates for cryopreservation is that the substrate according to the invention can be produced from a plurality of components (in particular anchoring axis, substrate plates) of identical or different plastics which ensure sufficient stability in all operating states and are movable relative to one another , It has surprisingly been found that the plastics used are relatively soft and deformable at room temperature, but nevertheless sufficiently stable. At the low preservation temperatures, however, the plastics are hard and inelastic, while maintaining their relative mobility with adapted thermal expansion coefficients.
Verfahrensbezogen beruht die Erfindung auf der allgemeinen technischen Lehre, Proben zur Kryokonservierung in einem erfindungsgemäßen Substrat mit einem Plattenstapel abzulegen und im Stapelverbund einzufrieren. Dabei kann die Bildung des Stapels vor oder nach der Ablage der Proben erfolgen. Die Beschickung des Substratplatten nach Bildung des Stapels kann den Vorteil besitzen, dass unbeabsichtigte Vertauschungen von Substratplatten vermieden werden. Die Beschickung des Substratplatten vor Bildung des Stapels kann Vorteile in Bezug auf die Handhabung der Substratplatten zum Beispiel in einem Labor besitzen. Gemäß einer vorteilhaften Variante der Erfindung werden einzelne Substratplatten im gefrorenen oder aufgetauten Zustand aus dem Stapelverbund geschwenkt und/oder geschoben, so dass einzelne Proben gezielt aus dem erfindungsgemäßen Substrat entnommen werden können.In terms of method, the invention is based on the general technical teaching of depositing samples for cryopreservation in a substrate according to the invention with a stack of plates and freezing them in the stacked composite. The formation of the stack can take place before or after the deposition of the samples. The loading of the substrate plates after formation of the stack may have the advantage that unintentional permutations of substrate plates are avoided. The loading of the substrate plates prior to formation of the stack may have advantages in handling the substrate plates, for example, in a laboratory. According to an advantageous variant of the invention, individual substrate plates in the frozen or thawed state are pivoted out of the stacked composite and / or pushed, so that individual samples can be removed in a targeted manner from the substrate according to the invention.
Weitere Einzelheiten und Vorteile der Erfindung werden aus der folgenden Beschreibung bevorzugter Ausführungsformen ersichtlich. Es zeigen:
- Figuren 1 bis 3:
- Perspektivansichten eines Drehstapel-Substrats gemäß der Erfindung;
- Figuren 4 und 5:
- Teilansichten von Substratplatten eines Drehstapel-Substrats von oben und von unten;
- Figur 6:
- eine Perspektivansicht einer Verankerungsachse eines Drehstapel-Substrats;
- Figuren 7 und 8:
- Illustrationen eines Basisteils eines Drehstapel-Substrats;
- Figur 9:
- eine Perspektivansicht eines Schiebestapel-Substrats gemäß der Erfindung;
- Figur 10:
- eine vergrößerte Darstellung eines Achsensegments;
- Figur 11:
- eine Teilansicht einer Substratplatte in einem Schiebestapel-Substrat; und
- Figur 12:
- ein Basisteil eines Schiebestapel-Substrats.
- FIGS. 1 to 3:
- Perspective views of a rotary stack substrate according to the invention;
- FIGS. 4 and 5:
- Partial views of substrate plates of a rotary stack substrate from above and below;
- FIG. 6:
- a perspective view of an anchoring axis of a rotary stack substrate;
- FIGS. 7 and 8:
- Illustrations of a base part of a rotary stack substrate;
- FIG. 9:
- a perspective view of a sliding stack substrate according to the invention;
- FIG. 10:
- an enlarged view of an axis segment;
- FIG. 11:
- a partial view of a substrate plate in a sliding stack substrate; and
- FIG. 12:
- a base part of a sliding stack substrate.
Die in den Figuren 1 bis 3 gezeigte bevorzugte Ausführungsform eines erfindungsgemäßen Drehstapel-Substrats 100 umfasst einen Stapel 10 aus Substratplatten 11, 12, 13, die mit einer Verankerungsachse 20 miteinander verbunden und auf einem Basisteil 60 angeordnet sind. Es kann vorgesehen sein, dass wenigstens eine Platte im Stapel 10 eine elektronische oder optische Datenspeichereinrichtung 50 (z. B. FLASH-Speicher) umfasst.The preferred embodiment of a
Die Substratplatten 11, 12, 13 sind jeweils ebene, plattenförmige Bauteile mit einer rechteckigen Grundform, auf deren oberen Seite die Kompartimentanordnung 40 mit einer Vielzahl von Probenreservoiren 41, 42, 43 gebildet ist. Die Probenreservoire 41, 42, 43 sind jeweils topfförmige Vertiefungen mit einem umlaufenden, kreisförmigen Rand. Die obere Seite der Substratplatten (z. B. 13 in Figur 2) weist einen umlaufenden Rand 17 auf, der sich an mindestens einer Seite zur Bereitstellung der Eingriffsmittel 30 (siehe unten) erweitert und der über die Plattenebene höher aufragt als die Ränder der Probenreservoire 41, 42, 43. Aus Schutzgründen kann über die obere Seite der Substratplatten eine Schutzfolie gespannt sein, die auf dem Rand 17 aufliegt.The
Die Substratplatten 11, 12, 13 sind aus Kunststoff oder ggf. einem Verbundmaterial aus einem Kunststoff hergestellt, in den ein Metall (zum Beispiel Aluminium) eingebettet ist. In mindestens eine der Substratplatten kann ein Speicher, wie zum Beispiel ein magnetischer, optischer oder elektronischer Speicher integriert (eingelegt, eingegossen oder eingespritzt) sein.The
Die Platten im Stapel 10 weisen an mindestens einer Seite Manipulationsöffnungen 70 (zum Beispiel 71 in Figur 2 oder 72 in Figur 9) auf. Die Manipulationsöffnungen 70 dienen dem Eingriff von Manipulationsgeräten, Werkzeugen oder anderen Hilfsgeräten, mit denen insbesondere der Transport des gesamten Substrats oder von einzelnen Platten durchgeführt wird.The plates in the
Die vergrößerte Teilansichten der Substratplatten 11, 12 in den Figuren 4 und 5 zeigen in einer Ecke der Substratplatte jeweils die Lagerbohrung 15, die sich über die Einführöffnung 16 zum Umfang der Substratplatten 11, 12 hin öffnet. Die Einführöffnung 16 besitzt eine sich etwa über die Hälfte der Dicke der Substratplatte 11 erstreckende Kragenöffnung 18, an der die Breite der durch die Einführöffnung 16 gebildeten Lücke geringer als der Durchmesser der Lagerbohrung 15 ist. Der Kragen (Rand der Kragenöffnung 18) bildet bei passender Ausrichtung der Verankerungsachse relativ zur Substratplatte ein Rückhaltelement (siehe unten).The enlarged partial views of the
Auf der unteren Seite der Substratplatte 11 (Figur 4) ist neben der Lagerbohrung 15 als Profilierung ein noppenförmiger Vorsprung 32 vorgesehen, der zusammen mit einer Profilierung, wie zum Beispiel der Ausnehmung 31 auf der angrenzenden oberen Seite der benachbarten Substratplatte 12 (Figur 5) die Eingriffsmittel 30 des Drehstapel-Substrats 100 bildet.On the lower side of the substrate plate 11 (FIG. 4), a knob-shaped
Die Verankerungsachse 20 umfasst gemäß Figur 6 einen durchgehenden Stab 21 mit einem bestimmten Außendurchmesser entsprechend dem Durchmesser der Lagerbohrungen 15 in den Substratplatten und einer am oberen Ende vorgesehenen Auskragung 22 mit einem größeren Durchmesser. Entlang der Länge des Stabes 21 sind Schnitt- oder Schlüsselflächen 23 vorgesehen, an denen die Dicke des Stabes 21 auf die Breite der Kragenöffnung 18 der Einführungsöffnung 16 vermindert ist. Die Schlüsselflächen 23 besitzen eine axiale Länge, die größer oder gleich der Länge der Kragenöffnungen 18 (in Stapelrichtung) sind, und einen axialen Abstand, der im wesentlichen dem Abstand der Kragenöffnungen 18 der Substratplatten in Stapelrichtung entspricht.According to FIG. 6, the anchoring
Am unteren Ende des Stabes 21 ist ein Gewinde 24 vorgesehen. Das Gewinde 24 kann über die gesamte Länge der Verankerungsachse 20 verlaufen, so dass diese wie eine Schraube aufgebaut ist. Diese Gestaltung ermöglicht einen einfachen Zuschnitt der gewünschten Länge einer Verankerungsachse. Am oberen Ende des Stabes 21 ist in der Auskragung 22 ein Schlitz 25 vorgesehen. Die Verankerungsachse 20 besteht aus Kunststoff oder ggf. einem Verbundmaterial aus Kunststoff, in den ein Metallkern (zum Beispiel aus Aluminium) eingebettet ist.At the lower end of the
Die in Figur 7 gezeigte Basisplatte 60 bildet einen untersten Träger für den Stapel 10 der Substratplatten 11, 12, 13. Die Basisplatte 60 besitzt auf ihrer oberen Seite eine Gewindebohrung 61, die entsprechend der Position der Lagerbohrungen 15 ausgerichtet und zur Aufnahme des Gewindes 24 der Verankerungsachse 20 eingerichtet ist. Des Weiteren ist analog zur Ausnehmung 31 eine Ausnehmung 62 auf der oberen Seite der Basisplatte 60 vorgesehen.The
Die Basisplatte 60 besitzt eine Ausnehmung 63 zur Aufnahme eines magnetischen, elektronischen oder optischen Datenspeichers (nicht dargestellt). Der Datenspeicher wird in die Ausnehmung 63 eingelegt und durch Vorsprünge 64 an den Rändern der Ausnehmung 63 fixiert. Alternativ kann ein Eingießen oder Einspritzen des Datenspeichers vorgesehen sein. Parallel zur Plattenebene besitzt die Ausnehmung 63 mindestens eine seitliche Öffnung, durch die einerseits auch bei zusammengesetztem Stapel der Datenspeicher einschiebbar ist und andererseits ein elektrischer Anschluss für den Datenspeicher durchführbar ist. So verweist das Bezugszeichen 65 auf eine Schnittstellenöffnung (siehe auch Figur 12). Wird der Datenspeicher beispielsweise durch einen Compact-FLASH-Speicher gebildet, kann durch die Schnittstellenöffnung 65 ein Stecker mit Kontaktstiften zum Anschluss an den Compact-FLASH-Speicher eingeführt und ggf. seitlich an der Basisplatte zumindest zeitweilig fixiert (z. B. mit einer Clips-Verbindung) werden. Über die Schnittstelle kann der Datenspeicher mit einer externen Steuereinrichtung verbunden werden.The
Die am Beispiel der Basisplatte 60 beschriebene Anbringung des Datenspeichers kann auch an mindestens einer der Substratplatten oder der Abdeckplatte vorgesehen sein.The attachment of the data memory described on the example of the
Figur 8 illustriert den ersten Schritt beim Aufbau eines erfindungsgemäßen Drehstapel-Substrats 100. Zunächst wird die Verankerungsachse 20 locker in das Basisteil 60 eingeschraubt, so dass die Schlüsselflächen 23 senkrecht zur Einführungsöffnung 16 stehen. In diesem angehobenen Zustand, der auch als Freigabeposition bezeichnet wird, befinden sich die Schlüsselflächen 23 entlang der Länge der Verankerungsachse jeweils in einer Höhe über der Basisplatte, dass die Kragenöffnungen (18) der Substratplatten im Stapel jeweils mit den Schlüsselflächen 23 ausgerichtet sind. In der Freigabeposition kann die Verankerungsachse durch die Einführungsöffnungen 16 in die Lagerbohrung eingeführt oder aus dieser heraus geschoben werden. In der Freigabeposition wird die unterste Substratplatte 11, die ggf. bereits mit Proben beschickt ist, auf die Basisplatte 60 geschoben. Da die unterste Schlüsselfläche 23 passend ausgerichtet ist, kann die Substratplatte 11 vorgeschoben werden, bis die Verankerungsachse 20 durch die Lagerbohrung 15 verläuft. Anschließend werden weitere Substratplatten entsprechend der Länge der verwendeten Verankerungsachse 20 aufgeschoben.FIG. 8 illustrates the first step in constructing a
Die Verankerungsachse 20 befindet sich nach Vervollständigung des Stapels 10 zunächst noch in dem angehobenen Zustand der Freigabeposition. Durch die Verdrehung der Verankerungsachse z. B. mit einem Schraubenzieher, der in den Schlitz 25 der Auskragung 22 (s. Figur 6) eingreift, wird die Verankerungsachse 20 abgesenkt. Wenn die Verankerungsachse 20 in die Basisplatte 60 eingeschraubt wird, geht die Ausrichtung der Schlüsselflächen 23 mit den Kragenöffnungen 18 entlang der Länge der Verankerungsachse 20 verloren. Die Substratplatten 11, 12, 13 können nicht mehr vom Stapel 10 getrennt werden. Beim Einschrauben wird zunächst ein Zustand erreicht, in dem die Substratplatten 11, 12, 13 zwischen der Auskragung 22 der Verankerungsachse 21 und der Basisplatte 60 in Stapelrichtung noch einen Spielraum besitzen und geringfügig beweglich sind. Dieser Zustand wird auch als Drehposition der Verankerungsachse 20 bezeichnet. In der Drehposition ist der Spielraum der Substratplatten 11, 12, 13 größer als die Höhe der Profilierungen 31, 32, so dass die Substratplatten 11, 12, 13 aus dem Stapel um die Verankerungsachse 20 herausgeschwenkt werden können.The anchoring
Zur Verriegelung des Stapelverbundes wird die Verankerungsachse 20 fest in die Basisplatte 60 eingeschraubt. Dieser Zustand wird auch als Fixierposition der Verankerungsachse 20 bezeichnet. In der Fixierposition werden die Substratplatten zusammengedrückt, so dass die Eingriffsmittel 31, 32 ineinander greifen und eine weitere Verschiebung oder Verschwenkung der Substratplatten blockieren.To lock the stack composite, the anchoring
In der Fixierposition kann das Einfrieren und Lagern des Substrats 100 zum Beispiel bei der Temperatur des flüssigen Stickstoffs oder im Dampf des flüssigen Stickstoffs (Normaldruck) erfolgen. Wenn einzelne Proben, wie z. B. aus der Substratplatte 12 gemäß Figur 1 entnommen werden sollen, kann die Verankerungsachse 20 durch Lockerung der Verschraubung an der Basisplatte 60 in die Drehposition überführt werden. In diesem Zustand werden die Eingriffsmittel 31, 32 freigegeben, so dass die Substratplatte 12 seitlich um die Verankerungsachse 20 nach außen verschwenkbar ist (Fig. 1). Die Verdrehung der Verankerungsachse 20 und/oder der Substratplatte 12 können so gewählt werden, dass die Schlüsselflächen 23 mit den Einführungsöffnungen 16 passend zusammenwirken, so dass die Substratplatte 12 vom Stapel 10 getrennt werden kann. Im Drehstapel-Substrat 100 sind bspw. 1 bis 20 Substratplatten übereinander angeordnet. Je nach der gewünschten Plattenzahl wird eine Verankerungsachse 20 mit geeigneter Länge verwendet.In the fixing position, the freezing and storage of the
In Figur 8 wird die Verankerungsachse zu Illustrationszwecken in der Fixierposition gezeigt, obwohl der Stapel noch nicht vervollständigt ist.In Figure 8, the anchoring axis is shown in the fixing position for illustrative purposes, although the stack is not yet completed.
Es stellt einen besonderen Vorteil der Erfindung dar, dass die Verankerungsachse zwischen den Freigabe-, Dreh- und Fixierpositionen allein durch eine Verdrehung, zum Beispiel durch das Einschrauben in die Basisplatte verstellt werden kann. Die Anhebung beim Schrauben wird durch die Steigung des Gewindes 24 bestimmt. Damit kann vorteilhafterweise durch die Zahl der Umdrehungen der Verankerungsachse die Überführung zwischen den verschiedenen Positionen festgelegt werden.It is a particular advantage of the invention that the anchoring axis between the release, rotational and fixing positions solely by a rotation, for example can be adjusted by screwing into the base plate. The increase in the screw is determined by the pitch of the
Allgemein ist an einem Substratstapel vorzugsweise mindestens ein Informationsträger vorgesehen, der durch die o. g. Datenspeicher und/oder durch zusätzliche Speichermedien, wie zum Beispiel einen Barcode gebildet wird.In general, at least one information carrier is preferably provided on a substrate stack, which is provided by the above-mentioned o. Data storage and / or by additional storage media, such as a bar code is formed.
Eine bevorzugte Ausführungsform eines Schiebestapel-Substrats 200 ist in den Figuren 9 bis 12 illustriert. Beim Schiebestapel-Substrat 200 sind die Eingriffsmittel 30 als formschlüssige Schiebeführung zum Beispiel aus Stegen 33 und Nuten 34 geformt, die als gerade Führungsschienen zueinander komplementär an den Rändern der Ober- und Unterseiten der Substratplatten gebildet sind. Im Stapel hintergreifen Stege 33 einer Substratplatte die Nuten 34 der benachbarten Substratplatte, so dass der Stapel durch aufeinanderfolgendes Aufschieben der Substratplatten gebildet wird. Auch bei dieser Variante kann eine einstückige Verankerungsachse 20 mit den Schlüsselflächen 23 vorgesehen sein, um nach Vervollständigung des Stapels 10 eine weitere Verschiebung der Substratplatten zu unterbinden.A preferred embodiment of a sliding
Bevorzugt wird jedoch bei dem Schiebestapel-Substrat eine aus Achsensegmenten 26 gebildete Verankerungsachse 20 verwendet. Diese besitzt den Vorteil, dass die Länge der Verankerungsachse 20 einfach durch die Zahl der benutzten Achsensegmente 26 passend zur Zahl der Substratplatten eingestellt wird.Preferably, however, an anchoring
Jedes Achsensegment 26 umfasst gemäß Figur 10 einen zylindrischen Körper, an dessen Ober- und Unterseiten zueinander komplementäre, schlitzförmige Ausnehmungen 27 und Vorsprünge 28 gebildet sind. Im zusammengesetzten Schiebestapel-Substrat 200 greift ein Vorsprung 28 jeweils in die Ausnehmung 27 des darunterliegenden Achsensegments 26. Wenn die Ausrichtung der schlitzförmigen Ausnehmungen und Vorsprünge 27, 28 parallel zur Ausrichtung der Eingriffsmittel 33, 34 verläuft, können Substratplatten durch Schieben in der Fügerichtung voneinander getrennt werden. Wenn die schlitzförmigen Ausnehmungen und Vorsprünge 27, 28 anders ausgerichtet sind, wird die gegenseitige Verschiebung der Substratplatten blockiert.Each
Die Achsensegmente 26 sind in den Lagerbohrungen 15 (siehe Figur 11) drehbar angeordnet. Vorzugsweise werden die Substratplatten mit den Achsensegmenten vorgefertigt. Bei der Vorfertigung werden die Achsensegmente bei Raumtemperatur in die Lagerbohrungen der Substratplatten eingepresst. Bei der Betriebstemperatur der Kryokonservierung, bei der die Elastizität der Materialien stark beschränkt ist, können die Achsensegmente 26 kaum mehr zerstörungsfrei aus den Lagerbohrungen 15 entfernt werden.The
Auch beim Schiebestapel-Substrat 200 ist ein Basisteil 60 (Figur 12) vorgesehen, in dem entsprechend ein Achsensegment 26 angeordnet ist.Also in the case of the sliding
Die Substratplatten und Verankerungsachsen der Substrate 100, 200 werden vorzugsweise durch Spritzguss aus TPX, PE, PTFE o. dgl. hergestellt. Die Seitenlängen der Substratplatten sind bspw. im Bereich von 10 mm bis 20 cm oder darüber, wie z. B. 50 cm oder 80 cm gewählt. Die Dicke der Substratplatten beträgt bspw. 4 mm bis 5 cm oder mehr. Die Zahl der Probenreservoire 41, 42, 43 je Substratplatte ist von der Größe der Substratplatte und der Probenreservoire abhängig und beträgt bspw. 20 bis 200 für kleinere Formate. Bei größeren Formaten kann die Zahl erheblich höher sein und zum Beispiel 5000 bis 10000 betragen.The substrate plates and anchoring axes of the
Die Größe und Form der Probenreservoire sind von den biologischen Proben (insbesondere biologisches Gewebe, Gewebeteile, biologische Zellen, Zellgruppen, Zellbestandteile, Zellorganellen oder biologisch relevante Makromoleküle) abhängig, die gelagert werden sollen.The size and shape of the sample reservoirs depends on the biological samples (in particular biological tissue, tissue parts, biological cells, cell groups, cell components, cell organelles or biologically relevant macromolecules) which are to be stored.
Abweichend von den dargestellten Ausführungsbeispielen können je nach den Anforderungen bei Anwendung des erfindungsgemäßen Substrats Modifizierungen insbesondere in Bezug auf die Geometrie der einzelnen Teile vorgesehen sein. Beispielsweise ist es erfindungsgemäß nicht zwingend erforderlich, dass alle Substratplatten die gleiche Grundfläche besitzen. Vielmehr können Substratplatten mit verschiedenen Grundflächen im Stapel kombiniert werden. Beispielsweise kann die Grundfläche im Stapel nach oben hin kleiner werden. Des Weiteren ist nicht zwingend vorgesehen, dass die Verankerungsachsen und Lagerbohrungen jeweils einen runden Querschnitt besitzen. Es kann auch eine Verankerungsachse mit einem eckigen Querschnitt vorgesehen sein. Schließlich können bei der Drehstapel-Variante die Schlüsselflächen relativ zueinander verschieden ausgerichtet sein, so dass bei der Verdrehung der Verankerungsachse jeweils eine Substratplatte freigegeben und die übrigen blockiert werden.Notwithstanding the illustrated embodiments may be provided depending on the requirements when using the substrate according to the invention modifications in particular with respect to the geometry of the individual parts. For example, it is not absolutely necessary according to the invention that all substrate plates have the same base area. Rather, substrate plates can be combined with different base areas in the stack. For example, the base in the stack can be smaller towards the top. Furthermore, it is not necessarily provided that the anchoring axes and bearing bores each have a round cross-section. It can also be provided an anchoring axis with a polygonal cross-section. Finally, in the rotary stacking variant, the key surfaces can be aligned differently relative to one another, so that in each case one substrate plate is released during the rotation of the anchoring axis and the others are blocked.
Die in der Beschreibung, den Zeichnungen und den Ansprüchen offenbarten Merkmale der Erfindung können einzeln oder in Kombination für die Verwirklichung der Erfindung in ihren verschiedenen Ausgestaltungen von Bedeutung sein. Insbesondere können die für das Drehstapel-Substrat beschriebenen Merkmale beim Schiebestapel-Substrat (oder umgekehrt) vorgesehen sein.The features of the invention disclosed in the specification, the drawings and the claims may, individually or in combination, be of importance for the realization of the invention in its various forms. In particular, the features described for the rotary stack substrate may be provided on the slide stack substrate (or vice versa).
Claims (27)
- Substrate (100, 200) for receiving and cryopreserving a multiplicity of samples, which comprises:- a multiplicity of substrate plates (11, 12, 13) which are disposed one above the other as a stack (10), and- an anchoring axle (20) with which the substrate plates (11, 12, 13) are connected,- each substrate plate (11, 12, 13) having a bearing boring (15) through which the anchoring axle (20) penetrates,characterised in that- the substrate plates (11, 12, 13) have respectively a compartment arrangement (40) with a multiplicity of sample reservoirs (41, 42, 43), and- at least one substrate plate (12) is pivotable about the anchoring axle (20) out of the stack (10).
- Substrate according to claim 1, in which the substrate plates (11, 12, 13) have a rectangular shape and the bearing boring (15) is provided respectively in one corner of the substrate plates (11, 12, 13).
- Substrate according to claim 1 or 2, in which the bearing boring (15) at least of one of the substrate plates (11, 12, 13) has at the edge an introduction opening (16) for lateral introduction of the anchoring axle (20) into the bearing boring (15).
- Substrate according to claim 3, in which the introduction opening (16) forms a collar opening (18) with a smaller width relative to the diameter of the bearing boring (15), and the anchoring axle (20) has a thickness, at least in partial portions of the length thereof, which is smaller than or the same as the width of the collar opening (18).
- Substrate according to at least one of the preceding claims in which the anchoring axle (20) has an overhang (22) at the upper end thereof.
- Substrate according to at least one of the preceding claims, in which the anchoring axle (20) is disposed rotatably.
- Substrate according to at least one of the preceding claims, in which the stack (10) contains at least one data logging device (50), a base plate (60) and/or a cover plate.
- Substrate according to claim 7, in which the base plate (60) contains a data logger (65).
- Substrate according to claim 7 or 8, in which the anchoring axle (20) is connected detachably to a lowermost substrate plate (11) or to the base plate (60).
- Substrate according to at least one of the preceding claims, in which at least one substrate plate (11, 12, 13) can be displaced in the stack (10) perpendicularly to the anchoring axle (20).
- Substrate according to at least one of the preceding claims, in which the substrate plates (11, 12, 13) have engagement means (30) which block lateral displacement of the substrate plates (11, 12, 13) at least in a direction perpendicular to a stacking direction.
- Substrate according to claim 11, in which the engagement means (30) include at least one profile (31) on a lateral face of a substrate plate (11, 12, 13) which cooperates with a complementary profile (32) on a lateral face of an adjacent substrate plate (11, 12, 13).
- Substrate according to claim 11 or 12, in which the anchoring axle (20) can be transferred, by rotation, from a lowered, fixing position, in which all the substrate plates (11, 12, 13) in the stack (10) are mutually fixed, into a rotating position, in which the substrate plates (11, 12, 13) can be moved corresponding to a clearance space in the stack direction and can be pivoted about the anchoring axle, and/or into a releasing position, in which at least one substrate plate (11, 12, 13) can be separated from the stack (10).
- Substrate according to claim 11, in which the engagement means (30) are formed by a form-fitting sliding guide.
- Substrate according to at least one of the preceding claims, in which the anchoring axle (20) includes a one-piece rod (21) which extends above the height of the stack (10).
- Substrate according to claims 15 and 4, in which the rod (21) has key faces (23) which form the partial portions with a thickness which is smaller than or the same as the width of the collar opening (18).
- Substrate according to at least one of the preceding claims 1 to 15, in which the anchoring axle (20) includes a multiplicity of axle segments (26).
- Substrate according to claim 17, in which the axle segments (26) have respectively a cylindrical body with a height which corresponds substantially to the thickness of the substrate plates (11, 12, 13) and with a diameter which corresponds to the diameter of the bearing borings (15), on upper and lower sides of the axle segments (26) recesses (27) and projections (28) being provided, which are complementary to each other and engage one in the other in the assembled stack (10) of substrate plates (11, 12, 13).
- Substrate according to at least one of the preceding claims, in which at least one substrate plate (11) contains a data logger.
- Substrate according to at least one of the preceding claims, in which the substrate plates are made of plastic material.
- Substrate according to at least one of the preceding claims, in which the substrate plates (11, 12, 13) have lateral lengths which are smaller than 10 cm.
- Method for cryopreserving samples, having a substrate according to at least one of the preceding claims, with the steps:- depositing the samples on the substrate plates (11, 12, 13), and- freezing the substrate plates (11, 12, 13) in the assembly of the stack (10).
- Method according to claim 22, in which the stack (10) of substrate plates (11, 12, 13) is produced before deposition of the samples.
- Method according to claim 22, in which the stack (10) of substrate plates (11, 12, 13) is produced after deposition of the samples.
- Method according to one of the claims 22 to 24, in which the individual substrate plates are pivoted and/or pushed out of the stack (10) in the frozen or thawed state.
- Use of a substrate according to at least one of the claims 1 to 21 for storing liquid or particulate samples.
- Use of a substrate according to at least one of the claims 1 to 22 for low temperature cryostorage of biological samples.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE10332296A DE10332296B3 (en) | 2003-07-16 | 2003-07-16 | Substrate stacks, in particular for the cryopreservation of biological samples |
PCT/EP2004/007956 WO2005007290A1 (en) | 2003-07-16 | 2004-07-16 | Stack of supports, in particular for cryopreservation of biological samples |
Publications (2)
Publication Number | Publication Date |
---|---|
EP1648607A1 EP1648607A1 (en) | 2006-04-26 |
EP1648607B1 true EP1648607B1 (en) | 2007-04-25 |
Family
ID=34071711
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP04741090A Not-in-force EP1648607B1 (en) | 2003-07-16 | 2004-07-16 | Stack of supports, in particular for cryopreservation of biological samples |
Country Status (6)
Country | Link |
---|---|
US (1) | US7608224B2 (en) |
EP (1) | EP1648607B1 (en) |
AT (1) | ATE360482T1 (en) |
BR (1) | BRPI0411988B1 (en) |
DE (2) | DE10332296B3 (en) |
WO (1) | WO2005007290A1 (en) |
Families Citing this family (14)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE102006055331B4 (en) * | 2006-01-27 | 2010-12-09 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Sample carriers and sample storage for the cryopreservation of biological samples |
DE102006059543B4 (en) * | 2006-12-16 | 2015-02-05 | Institut für Luft- und Kältetechnik gGmbH | Device for receiving and fixing tissue engineered constructs |
DE102008016217A1 (en) | 2008-03-28 | 2009-10-01 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Method and device for producing frozen biological particles |
ATE512894T1 (en) * | 2008-10-08 | 2011-07-15 | Zeta Biopharma Gmbh | DEVICE FOR RECEIVING CONTAINERS AND ASSOCIATED CONTAINERS |
US9086347B2 (en) * | 2011-06-03 | 2015-07-21 | Provia Laboratories, LLC | Cryopreservation device with retrofitted electronic tracking device |
US20130180171A1 (en) * | 2012-01-13 | 2013-07-18 | Kevin Oldenburg | Systems and Methods for Harvesting and/or Analyzing Biological Samples |
US9664431B2 (en) * | 2012-03-12 | 2017-05-30 | The World Egg Bank, Inc | Cryogenic sample holder |
DE102012013267A1 (en) * | 2012-07-04 | 2014-01-09 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Substrate device, preservative device and method for cryopreserving a biological sample |
US10690574B2 (en) * | 2016-05-11 | 2020-06-23 | The Florida International University Board Of Trustees | Chilling apparatus |
GB2548434B (en) * | 2016-09-27 | 2018-04-18 | Planer Plc | Incubator stand and incubator docking system |
MX369254B (en) * | 2017-03-06 | 2019-09-24 | Centro De Investig Cientifica Y De Educacion Superior De Ensenada Baja California Cicese | System for handling high-viscosity biological samples. |
CN110817069A (en) * | 2019-11-28 | 2020-02-21 | 湖南德凌物流配送有限公司 | Intelligent cryopreservation frame |
CN111389478B (en) * | 2020-03-23 | 2023-07-25 | 西安医学院 | Experimental treatment device for tissue slices and application method thereof |
CN115161179B (en) * | 2022-06-29 | 2023-05-19 | 生态环境部南京环境科学研究所 | Device and method for testing ecological toxicity of solid waste leachate based on zebra fish embryo |
Family Cites Families (15)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4761378A (en) * | 1983-03-04 | 1988-08-02 | American Home Products Corp. (Del.) | Microbiological testing apparatus |
US4676951A (en) * | 1985-07-01 | 1987-06-30 | American Hospital Supply Corp. | Automatic specimen analyzing system |
US5233844A (en) | 1991-08-15 | 1993-08-10 | Cryo-Cell International, Inc. | Storage apparatus, particularly with automatic insertion and retrieval |
DE4300231C1 (en) * | 1993-01-07 | 1993-12-02 | Schulz Peter | Petri dish - has side mounting with upper projecting pin and lower recess to form stack with swing access to each dish |
DE19752085A1 (en) * | 1996-12-21 | 1998-06-25 | Evotec Biosystems Gmbh | Polycarbonate carrier disc for large numbers of samples |
EP0904841B1 (en) * | 1997-09-29 | 2004-01-28 | F. Hoffmann-La Roche Ag | Compound handling system comprising racks and containers |
DE10002666A1 (en) * | 2000-01-21 | 2001-08-02 | Greiner Bio One Gmbh | Containers for the storage of biological material |
US6455007B1 (en) * | 2000-06-13 | 2002-09-24 | Symyx Technologies, Inc. | Apparatus and method for testing compositions in contact with a porous medium |
US6699437B1 (en) * | 2000-08-29 | 2004-03-02 | Thomas W. Astle | Bioassay cassette with memory and method of use |
EP1320709A1 (en) | 2000-09-07 | 2003-06-25 | Jin-Hee Kim | Storage device of kimchi refrigerator |
US20020051995A1 (en) * | 2000-10-30 | 2002-05-02 | Rajan Kumar | Stacked arrays |
US6446807B1 (en) * | 2000-11-24 | 2002-09-10 | 3088081 Canada, Inc. | Assembly of modular containers for handling, transporting and storing microscope specimen slides |
DE10144925A1 (en) * | 2001-09-12 | 2003-03-27 | Fraunhofer Ges Forschung | Cryopreservation method for cells and cell components comprises applying samples to e.g. a CD as a storage substrate and burning position-specific data on to disk. |
EP1340062B1 (en) | 2000-12-07 | 2012-10-24 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Cryostorage method and device |
WO2002087760A1 (en) * | 2001-04-30 | 2002-11-07 | Epr Labautomation Ag | Method and device for storing and dosing small quantities of liquid |
-
2003
- 2003-07-16 DE DE10332296A patent/DE10332296B3/en not_active Expired - Fee Related
-
2004
- 2004-07-16 WO PCT/EP2004/007956 patent/WO2005007290A1/en active IP Right Grant
- 2004-07-16 US US10/564,491 patent/US7608224B2/en not_active Expired - Fee Related
- 2004-07-16 DE DE502004003628T patent/DE502004003628D1/en active Active
- 2004-07-16 AT AT04741090T patent/ATE360482T1/en active
- 2004-07-16 EP EP04741090A patent/EP1648607B1/en not_active Not-in-force
- 2004-07-16 BR BRPI0411988-6A patent/BRPI0411988B1/en not_active IP Right Cessation
Also Published As
Publication number | Publication date |
---|---|
DE502004003628D1 (en) | 2007-06-06 |
EP1648607A1 (en) | 2006-04-26 |
BRPI0411988A (en) | 2006-10-17 |
US7608224B2 (en) | 2009-10-27 |
ATE360482T1 (en) | 2007-05-15 |
BRPI0411988B1 (en) | 2015-02-24 |
DE10332296B3 (en) | 2005-04-28 |
WO2005007290A1 (en) | 2005-01-27 |
US20060154232A1 (en) | 2006-07-13 |
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